CN114689853A - Mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibody detection test strip, combined detection card, method and application thereof - Google Patents
Mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibody detection test strip, combined detection card, method and application thereof Download PDFInfo
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Abstract
The invention discloses a mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibody detection test strip, belonging to the technical field of in vitro diagnostic reagents. This test paper strip includes the sample pad, the gold mark pad, the nitrocellulose membrane, absorbent paper, the bottom plate, the upper portion laminating of bottom plate has the nitrocellulose membrane, the upper portion both ends of nitrocellulose membrane have laminated the gold mark pad and absorbent paper respectively, the laminating of gold mark pad upper portion one end has the sample pad, the recombinant antigen of the corresponding pathogen of fixed colloidal gold mark's mouse IgG and colloidal gold mark is stamped to the gold mark, the nitrocellulose membrane is equipped with the quality control line of peridium mouse anti human IgM monoclonal antibody's detection line and peridium goat anti mouse IgG antibody in proper order. The test strip can realize in-vitro qualitative detection of mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and coxsackie virus group B IgM antibodies in human whole blood or serum samples, and has the advantages of high sensitivity, high specificity and short time consumption.
Description
Technical Field
The invention belongs to the technical field of in-vitro diagnostic reagents, and particularly relates to a mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibody detection test strip, a combined detection card, a method and an application thereof.
Background
Mycoplasma Pneumoniae (MP) is one of the common pathogens causing upper respiratory tract infections; chlamydia Pneumoniae (CP) is an important pathogen of human respiratory diseases and can cause acute and chronic respiratory diseases, and 5-10% of community-acquired pneumonia, bronchitis and nasosinusitis are caused by the Chlamydia pneumoniae; respiratory Syncytial Virus (RSV) is a enveloped mononegavirale RNA virus belonging to the genus pneumoniaceae of the family paramyxoviridae, the most common Respiratory infectious pathogen that seriously harms infant health worldwide; adenovirus (ADV), one of the major pathogens of viral pneumonia, is a DNA virus that propagates primarily within cells; coxsackievirus group B (CoxB) is a common virus infecting human body through respiratory tract and digestive tract, and can cause respiratory tract infection, diarrhea, meningitis, myocarditis, serious infection of newborn and the like; the existing detection methods of the mycoplasma pneumoniae/chlamydia pneumoniae/respiratory syncytial virus/adenovirus/coxsackie virus B group are approximately the same, the culture method has high sensitivity but much time consumption, and the requirement on the biological safety level of a laboratory is high; although molecular biology techniques are both sensitive and specific and are time consuming, they require expensive equipment and specialized technicians, which have significant limitations for clinical implementation.
Disclosure of Invention
In order to overcome the above disadvantages in the prior art, one of the purposes of the present invention is to provide a test strip for detecting IgM antibodies against mycoplasma pneumoniae, chlamydia pneumoniae and respiratory viruses, which can realize in vitro qualitative detection of the IgM antibodies against mycoplasma pneumoniae/chlamydia pneumoniae/respiratory syncytial virus/adenovirus/coxsackie virus group B in human whole blood or serum samples, and has the advantages of high sensitivity, high specificity and short time consumption.
The invention also aims to provide a combined detection card for mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibodies.
The invention also aims to provide a combined detection kit for mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibodies.
The fourth purpose of the invention is to provide a preparation method of the combined detection card for mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibody.
The fifth purpose of the invention is to provide a detection method of mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibody, which can complete the detection of five pathogens simultaneously under the condition of only one sampling, greatly facilitates patients and clinical inspectors, and has better specificity and sensitivity.
In order to achieve one of the purposes, the invention adopts the following technical scheme:
the invention provides a mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibody detection test strip, which comprises a sample pad, a gold mark pad, a nitrocellulose membrane, absorbent paper and a bottom plate, wherein the nitrocellulose membrane is attached to the upper part of the bottom plate, the gold mark pad and the absorbent paper are respectively attached to two ends of the upper part of the nitrocellulose membrane, the sample pad is attached to one end of the upper part of the gold mark pad, and the test strip is characterized in that colloidal gold-labeled mouse IgG and colloidal gold-labeled mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus or coxsackie virus B group recombinant antigen are fixed on the gold mark pad, and the nitrocellulose membrane is sequentially provided with a detection line coated with mouse anti-human IgM monoclonal antibody and a quality control line coated with goat anti-mouse IgG.
In order to achieve the second purpose, the invention adopts the following technical scheme:
the invention also provides a combined detection card for the mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibodies, which comprises a plastic shell provided with the detection test paper strip for the mycoplasma pneumoniae, the chlamydia pneumoniae, the respiratory syncytial virus, the adenovirus and the Coxsackie group B virus antibodies, wherein the plastic shell is provided with five groups of sample adding holes and an observation window, the sample adding holes are arranged above the sample pad, and the observation window is arranged above the detection line and the quality control line.
In order to achieve the third purpose, the invention adopts the following technical scheme:
the invention also provides a combined detection kit for the IgM antibodies of the mycoplasma pneumoniae, the chlamydia pneumoniae and the respiratory viruses, which comprises a combined detection card for the IgM antibodies of the mycoplasma pneumoniae, the chlamydia pneumoniae, the respiratory syncytial viruses, the adenoviruses and the coxsackie viruses in the B group.
In order to achieve the fourth purpose, the invention adopts the following technical scheme:
the invention also provides a preparation method of the combined detection card for mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibodies, which comprises the following specific steps:
s1, labeling the mouse IgG monoclonal antibody with colloidal gold to obtain the mouse IgG labeled with the colloidal gold; respectively labeling the recombinant antigens of the group B of mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and coxsackie virus by using the colloidal gold to obtain corresponding antigen-colloidal gold compounds; respectively mixing the mouse IgG labeled by the colloidal gold and the corresponding antigen-colloidal gold compound, and spraying the mixture on the gold label pad;
s2, coating the mouse anti-human IgM monoclonal antibody on a nitrocellulose membrane to form a detection line; coating goat anti-mouse IgG on the nitrocellulose membrane positioned on the other side of the detection line respectively to form a quality control line; drying to prepare the nitrocellulose membrane provided with the detection line and the quality control line;
s3, sequentially overlapping the corresponding sample pad, the gold mark pad, the nitrocellulose membrane and the absorbent paper on a bottom plate respectively to form a corresponding plate, and cutting the plate into a detection test strip for mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and coxsackie virus group B IgM antibody;
s4, putting the prepared Mycoplasma pneumoniae, Chlamydia pneumoniae, respiratory syncytial virus, adenovirus and Coxsackie virus group B IgM antibody detection test strips into the plastic shell, adjusting the positions of the Mycoplasma pneumoniae, Chlamydia pneumoniae, respiratory syncytial virus, adenovirus and Coxsackie virus group B IgM antibody detection test strips, enabling the sample adding hole to be above the sample pad, and enabling the observation window to be above the detection line and the quality control line, so as to obtain the Mycoplasma pneumoniae, Chlamydia pneumoniae, respiratory syncytial virus, adenovirus and Coxsackie virus group B IgM antibody combined detection card.
Further, in the S1, the drying temperature is 45-55 ℃, the relative humidity is less than or equal to 20%, and the drying time is 0.5-2 h; in the S2, the drying temperature is 55-60 ℃, the relative humidity is less than or equal to 20%, and the drying time is 14-16 h.
In order to realize the fifth purpose, the invention adopts the following technical scheme:
the invention also provides a detection method of the IgM antibodies of the mycoplasma pneumoniae, the chlamydia pneumoniae and the respiratory viruses, wherein the detection method is used for detecting a sample to be detected by using the combined detection card of the IgM antibodies of the mycoplasma pneumoniae, the chlamydia pneumoniae, the respiratory syncytial viruses, the adenoviruses and the coxsackie viruses in the B group, and comprises the following steps of:
s1, dripping the sample to be detected from whole blood or the sample to be detected from serum into the five groups of sample adding holes of the combined detection card for the mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and coxsackie virus group B IgM antibodies in sequence;
s2, sequentially dripping the sample diluent into the five groups of sample adding holes, standing, and observing and interpreting results;
s3, interpretation of results: if the sample liquid contains IgM antibodies of mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and/or coxsackie virus B group, presenting corresponding color bands on the detection line position of the corresponding test strip; if the sample to be detected does not contain IgM antibodies of mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus or coxsackie virus group B, the color is not developed on the detection line of the corresponding test strip; and no matter whether the sample liquid contains IgM antibodies of mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and/or coxsackie virus B group, the quality control line of each detection strip should present a color strip, otherwise, the detection is invalid.
Further, in S1, 10 μ L of the whole blood sample to be tested is sequentially added to the five groups of wells of the combined detection card for mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus, and coxsackie virus group B IgM antibodies, or 5 μ L of the serum sample to be tested is sequentially added to the five groups of wells of the combined detection card for mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus, and coxsackie virus group B IgM antibodies.
Further, in S2, 90 μ L of the sample dilution is sequentially dropped into the five groups of wells.
Further, in the step S2, the result is interpreted after the mixture is left to stand for 15 to 25 min.
The detection principle of the invention is as follows:
when a positive sample is detected, the Mycoplasma pneumoniae/Chlamydia pneumoniae/respiratory syncytial virus/adenovirus/Coxsackie virus group B IgM antibodies contained in the sample can be respectively combined with colloidal gold labeled Mycoplasma pneumoniae, Chlamydia pneumoniae, respiratory syncytial virus, adenovirus and Coxsackie virus group B recombinant antigens on a gold label pad to form a compound, and the compound is captured by mouse anti-human IgM at a detection line so as to agglutinate and develop color; the colloidal gold labeled mouse IgG is combined with the goat anti-mouse IgG coated on the quality control line for agglutination and color development. When a negative sample is detected, the sample does not contain mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and coxsackie virus group B IgM antibody, cannot form an immune complex, and only can develop color at a quality control line. Therefore, a red band appears at the control line (C) regardless of the presence of Mycoplasma pneumoniae, Chlamydia pneumoniae, respiratory syncytial virus, adenovirus, and coxsackie virus group B IgM antibodies in the clinical specimen.
Compared with the prior art, the invention has the following beneficial effects:
(1) the combined detection kit for the mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibodies provided by the invention can qualitatively detect the mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and coxsackie virus group B IgM antibodies in a human whole blood or serum sample in vitro by utilizing an immunochromatography technology and adopting a combined detection mode, and has the advantages of high sensitivity and short time consumption.
(2) The invention provides a detection method of mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibodies, which is a rapid immunology method, and is characterized in that symptoms of mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and coxsackie virus B group after infection are similar and are often infected, and specific IgM antibodies can be generated and released into body fluids such as serum, so that the five-unit detection kit is adopted, the detection of five pathogens is completed simultaneously under the condition of only one sampling, the detection method is greatly convenient for patients and clinical inspectors, and the specificity and the sensitivity are good.
(3) The preparation method of the combined detection card for mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibodies provided by the invention is simple, low in cost and easy for commercial production.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the embodiments or the prior art descriptions will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
Fig. 1 is a schematic side view of a test strip provided in embodiment 1 of the present application.
Fig. 2 is a schematic top view of a detection card provided in embodiment 2 of the present application.
Fig. 3 is a schematic view illustrating interpretation of a detection result of the detection card provided in embodiment 5 of the present application.
In the figure: 1. a sample pad; 2. a gold label pad; 3. a nitrocellulose membrane; 31. detecting lines; 32. a quality control line; 4. absorbent paper; 5. a base plate; 6. a sample application hole; 7. and (4) an observation window.
Detailed Description
In order to make the technical problems, technical solutions and beneficial effects to be solved by the present application clearer and more clearly apparent, the technical solutions of the present application will be clearly and completely described below with reference to the embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the present application and are not intended to limit the present application.
The reagents or instruments used are not indicated by the manufacturer, and are conventional products available commercially.
Example 1
The embodiment provides a mycoplasma pneumoniae, chlamydia pneumoniae, respiratory virus IgM antibody detection test strip, please refer to fig. 1, the test strip includes a sample pad 1, a gold-labeled pad 2, a nitrocellulose membrane 3, absorbent paper 4, and a bottom plate 5, the nitrocellulose membrane 3 is attached to the upper portion of the bottom plate 5, the gold-labeled pad 2 and the absorbent paper 4 are respectively attached to two ends of the upper portion of the nitrocellulose membrane 3, the sample pad 1 is attached to one end of the upper portion of the gold-labeled pad 2, a colloidal gold-labeled mouse IgG and a colloidal gold-labeled mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus or coxsackie virus B-group recombinant antigen are fixed on the gold-labeled pad 2, and the nitrocellulose membrane 3 is sequentially provided with a detection line 31 coated with a mouse IgM monoclonal antibody and a quality control line 32 coated with a goat anti-human IgG.
Example 2
The embodiment provides a mycoplasma pneumoniae, chlamydia pneumoniae, respiratory virus IgM antibody combined detection card, please refer to fig. 2, the detection card comprises a plastic shell containing the mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus, coxsackie group B IgM antibody detection test strip provided in embodiment 1, wherein the plastic shell is provided with a sample adding hole 6 and an observation window 7, the sample adding hole 6 is above a sample pad 1, and the observation window 7 is above a detection line 31 and a quality control line 32.
Example 3
The embodiment provides a combined detection kit for mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibodies, and referring to fig. 2, the combined detection card for mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and coxsackie virus group B IgM antibodies, the sample diluent and the instruction manual provided in the embodiment 2 are boxed to obtain the kit.
Example 4
The embodiment provides a preparation method of a mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibody combined detection card, which comprises the following specific steps:
1) labeling a mouse IgG monoclonal antibody by using colloidal gold to obtain a mouse IgG labeled by the colloidal gold; respectively marking recombinant antigens of the B groups of mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and coxsackie virus by using colloidal gold to obtain corresponding antigen-colloidal gold compounds; respectively mixing the mouse IgG marked by the colloidal gold and the corresponding antigen-colloidal gold compound, respectively spraying the mixture on the gold-labeled pads 2, and drying to form corresponding gold-labeled pads 2; wherein the drying temperature is 45-55 ℃, the relative humidity is less than or equal to 20 percent, and the drying time is 0.5-2 h;
2) coating a mouse anti-human IgM monoclonal antibody on a nitrocellulose membrane 3 to form a detection line 31; coating goat anti-mouse IgG on the cellulose nitrate membrane 3 at the other side of the detection line 31 respectively to form a quality control line 32; drying to prepare the nitrocellulose membrane 3 provided with a detection line 31 and a quality control line 32; wherein the drying temperature is 55-60 ℃, the relative humidity is less than or equal to 20 percent, and the drying time is 14-16 h.
3) The corresponding sample pad 1, the gold label pad 2, the nitrocellulose membrane 3 and the absorbent paper 4 are respectively overlapped on the bottom plate 5 in sequence to form a corresponding plate, and then the plate is cut into corresponding test strips;
4) assembling the prepared Mycoplasma pneumoniae, Chlamydia pneumoniae, respiratory syncytial virus, adenovirus and Coxsackie virus group B virus IgM antibody detection strips respectively to obtain a Mycoplasma pneumoniae, Chlamydia pneumoniae, respiratory syncytial virus, adenovirus and Coxsackie virus group B IgM antibody combined detection card;
5) when the kit is assembled, except for a certain number of detection cards sealed by aluminum foil bags, a corresponding amount of sample diluent and an instruction are put into the kit to complete the assembly.
Example 5
The embodiment provides a detection method of mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibodies, which comprises the following specific steps:
(1) sample processing
1. The whole blood or serum sample is collected according to a conventional method, and aseptic operation is ensured in the sample collection and preservation process.
2. The whole blood sample can be anticoagulated by heparin (9.8-28 IU/mL), sodium citrate (3.8%) and ethylenediamine tetraacetic acid (4.55mmoL/mL +/-0.85 mmoL/mL) in conventional amounts.
3. The whole blood sample can not be frozen and stored at the temperature of 2-8 ℃ for 3 days, the sample is best detected after being collected on the same day, and the diluted sample is not suitable for storage; the serum sample is not repeatedly frozen and thawed to prevent the antibody titer from being reduced, the freezing and thawing times cannot be more than 6, and the serum sample can be stored for 7 days at the temperature of 2-8 ℃, can be stored for 2 years at the temperature of-20 ℃ and can be stored for 5 years at the temperature of-70 ℃.
4. During detection, the sample is melted and mixed evenly, and the turbid or precipitated sample is detected after centrifugation or filtration clarification.
(2) Method of using kit
1. Before use, the kit and the specimen are placed at room temperature for rewarming for at least half an hour to ensure that the temperature is restored to the room temperature;
2. taking out the detection card from the aluminum foil bag, compiling a sample number, and placing the sample number on a horizontal desktop, wherein the detection card is shown as an attached figure 2;
3. adding 10 mu L of whole blood to be detected or 5 mu L of serum to be detected into each hole of the detection card sample adding hole 6, and then adding 90 mu L of sample diluent;
4. after 15min, the results were read within 25 min.
(3) Positive judgment value
The positive judgment value of the mycoplasma pneumoniae IgM antibody is not less than 1.2(S/C value), the positive judgment value of the chlamydia pneumoniae IgM antibody is not less than 1.2(S/C value), the positive judgment value of the respiratory syncytial virus IgM antibody is not less than 1.2(S/C value), the positive judgment value of the adenovirus IgM antibody is not less than 1.2(S/C value), the positive judgment value of the Coxsackie virus B group IgM antibody is not less than 1.2(S/C value), and according to the difference of regions and the difference of crowds, each laboratory is recommended to establish a reference range.
(4) Result judgment
Please refer to fig. 3
a. Negative: only one purple-red band appears at the quality control line 32(C), and no purple-red band appears at the detection line 31 (T). Negative results indicate that: the specimen does not contain Mycoplasma pneumoniae/Chlamydia pneumoniae/respiratory syncytial virus/adenovirus/coxsackie virus group B IgM antibody or the content thereof is lower than a detectable range.
b. Positive: one purple-red band appears at the control line 32(C) and the other purple-red band appears at the detection line 31 (T). The positive result shows that: the specimen contains mycoplasma pneumoniae/chlamydia pneumoniae/respiratory syncytial virus/adenovirus/coxsackie virus group B IgM antibody.
c. And (4) invalidation: the absence of a magenta band at the control line 32(C) indicates an incorrect procedure or a damaged test card.
Examples of the experiments
Clinical Performance evaluation test of the kit of the present invention
Serum samples of 50 patients infected with mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus, coxsackie virus group B virus and 50 normal persons were tested using the combined detection kit for mycoplasma pneumoniae, chlamydia pneumoniae, respiratory virus IgM antibodies and the common colloidal gold method kit provided in example 3, and the results are shown in table 1:
TABLE 1 comparison of detection sensitivity of the kit of the present invention and the kit of the conventional colloidal gold method
As can be seen from the table, the serum specimen detection by using the combined detection kit for mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibodies is excellent in specificity, and the detection sensitivity is high and reaches 96% compared with that of a common colloidal gold method kit.
The above embodiments are only preferred embodiments of the present invention, and the protection scope of the present invention is not limited thereby, and any insubstantial changes and substitutions made by those skilled in the art based on the present invention are within the protection scope of the present invention.
Claims (10)
1. The utility model provides a mycoplasma pneumoniae, chlamydia pneumoniae, respiratory virus IgM antibody detection test paper strip, includes sample pad, gold mark pad, nitrocellulose membrane, absorbent paper, bottom plate, the upper portion laminating of bottom plate has nitrocellulose membrane, nitrocellulose membrane upper portion both ends have been laminated respectively gold mark pad with absorbent paper, gold mark pad upper portion one end laminating has sample pad, its characterized in that, the gold mark is filled up the mouse IgG of fixed colloidal gold mark and the recombinant antigen of colloidal gold mark mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus or coxsackie virus B group, nitrocellulose membrane is equipped with the detection line of peridium mouse anti human IgM monoclonal antibody and the quality control line of peridium goat anti mouse IgG in proper order.
2. A combined detection card for mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibodies, which comprises a plastic shell provided with the test strip of claim 1, wherein the plastic shell is provided with five groups of sample adding holes and an observation window, the sample adding holes are arranged above a sample pad, and the observation window is arranged above the detection line and the quality control line.
3. A combined detection kit for IgM antibodies of mycoplasma pneumoniae, chlamydia pneumoniae and respiratory viruses, which is characterized by comprising the combined detection card for IgM antibodies of mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial viruses, adenoviruses and coxsackie viruses B group in claim 2.
4. The Mycoplasma pneumoniae, Chlamydia pneumoniae, respiratory virus IgM antibody combined detection kit according to claim 3, characterized by further comprising a sample diluent, wherein the sample diluent is a phosphate buffer.
5. The method for preparing the mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibody combined detection card according to claim 2, which is characterized by comprising the following steps:
s1, labeling the mouse IgG monoclonal antibody with colloidal gold to obtain the mouse IgG labeled with the colloidal gold; respectively labeling the recombinant antigens of the B groups of mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and coxsackie virus by using the colloidal gold to obtain corresponding antigen-colloidal gold compounds; respectively mixing the mouse IgG labeled by the colloidal gold and the corresponding antigen-colloidal gold compound, and spraying the mixture on the gold label pad;
s2, coating the mouse anti-human IgM monoclonal antibody on a nitrocellulose membrane to form a detection line; coating goat anti-mouse IgG on the nitrocellulose membrane positioned on the other side of the detection line respectively to form a quality control line; drying to prepare the nitrocellulose membrane provided with the detection line and the quality control line;
s3, sequentially overlapping the corresponding sample pad, the gold mark pad, the nitrocellulose membrane and the absorbent paper on a bottom plate respectively to form a corresponding plate, and cutting the plate into a detection test strip for mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and coxsackie virus group B IgM antibody;
s4, putting the prepared Mycoplasma pneumoniae, Chlamydia pneumoniae, respiratory syncytial virus, adenovirus and Coxsackie virus group B IgM antibody detection test strips into the plastic shell, adjusting the positions of the Mycoplasma pneumoniae, Chlamydia pneumoniae, respiratory syncytial virus, adenovirus and Coxsackie virus group B IgM antibody detection test strips, enabling the sample adding hole to be above the sample pad, and enabling the observation window to be above the detection line and the quality control line, so as to obtain the Mycoplasma pneumoniae, Chlamydia pneumoniae, respiratory syncytial virus, adenovirus and Coxsackie virus group B IgM antibody combined detection card.
6. The method for preparing the combined detection card for the mycoplasma pneumoniae, chlamydia pneumoniae and respiratory virus IgM antibodies according to claim 5, wherein in S1, the drying temperature is 45-55 ℃, the relative humidity is less than or equal to 20%, and the drying time is 0.5-2 h; in the S2, the drying temperature is 55-60 ℃, the relative humidity is less than or equal to 20%, and the drying time is 14-16 h.
7. A detection method of IgM antibodies of mycoplasma pneumoniae, chlamydia pneumoniae and respiratory viruses is characterized in that a sample to be detected is detected by the combined detection card of IgM antibodies of mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and coxsackie virus B group according to claim 2, and the detection method comprises the following steps:
s1, dripping the sample to be detected from whole blood or the sample to be detected from serum into the five groups of sample adding holes of the combined detection card for the mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and coxsackie virus group B IgM antibodies in sequence;
s2, sequentially dripping the sample diluent into the five groups of sample adding holes, standing, and observing and interpreting results;
s3, interpretation of results: if the sample liquid contains IgM antibodies of mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and/or coxsackie virus B group, presenting corresponding color bands on the detection line position of the corresponding test strip; if the sample to be detected does not contain IgM antibodies of mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus or coxsackie virus B group, the color is not developed on the detection line of the corresponding test strip; and no matter whether the sample liquid contains IgM antibodies of mycoplasma pneumoniae, chlamydia pneumoniae, respiratory syncytial virus, adenovirus and/or coxsackie virus B group, the quality control line of each detection strip should present a color strip, otherwise, the detection is invalid.
8. The method according to claim 7, wherein in step S1, 10 μ L of the whole blood sample is sequentially added to the five groups of wells of the Mycoplasma pneumoniae, Chlamydia pneumoniae, respiratory syncytial virus, adenovirus, Coxsackie virus group B IgM antibody combined test card or 5 μ L of the serum sample is sequentially added to the five groups of wells of the Mycoplasma pneumoniae, Chlamydia pneumoniae, respiratory syncytial virus, adenovirus, Coxsackie virus group B IgM antibody combined test card.
9. The method according to claim 7, wherein in S2, 90 μ L of the diluted sample solution is sequentially added dropwise to the five groups of wells.
10. The method for detecting Mycoplasma pneumoniae, Chlamydia pneumoniae, and respiratory virus IgM antibodies according to claim 7, wherein the result is interpreted after leaving it at S2 for 15 to 25 min.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117471098A (en) * | 2023-12-27 | 2024-01-30 | 北京芯迈微生物技术有限公司 | Mycoplasma pneumoniae and Chlamydia pneumoniae IgM antibody combined detection chip and application |
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2022
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117471098A (en) * | 2023-12-27 | 2024-01-30 | 北京芯迈微生物技术有限公司 | Mycoplasma pneumoniae and Chlamydia pneumoniae IgM antibody combined detection chip and application |
| CN117471098B (en) * | 2023-12-27 | 2024-03-12 | 北京芯迈微生物技术有限公司 | Mycoplasma pneumoniae and Chlamydia pneumoniae IgM antibody combined detection chip and application |
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